Morphological profiling of human T and NK lymphocytes by high-content cell imaging

The immunological synapse is a complex structure that decodes stimulatory signals into adapted lymphocyte responses. As such, it represents a unique window to monitor lymphocyte activity, owing to the development of systematic quantitative approaches applicable to primary cells. Here we demonstrate the applicability of high-content imaging to human T and NK cells and develop a pipeline for unbiased analysis of high-definition morphological profiles. Our approach reveals how distinct facets of actin cytoskeleton remodeling shape immunological synapse architecture and impact lytic granule positioning. Morphological profiling of CD8+ T cells from immunodeficient patients allows discriminating the roles of the ARP2/3 subunit ARPC1B and the ARP2/3 activator WASP in immunological synapse assembly. Single cell analysis further identifies uncoupling of lytic granules and F-actin radial distribution in ARPC1B-deficient lymphocytes. Our study provides a foundation for the development of morphological profiling as a scalable approach to monitor primary lymphocyte responsiveness and to identify complex aspects of lymphocyte micro-architecture.

German Y, Vulliard L, Kamnev A, Pfajfer L, Huemer J, Mautner AK, Rubio A, Kalinichenko A, Boztug K, Ferrand A, Menche J, Dupré L

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